Abstract
Although protein dynamics has been recognized as a potentially important contributor to enzyme catalysis, structural disorder is generally considered to reduce catalytic efficiency. This widely held assumption has recently been challenged by the finding that an engineered chorismate mutase combines high catalytic activity with the properties of a molten globule, a loosely packed and highly dynamic conformational ensemble. Taking advantage of the ordering observed upon ligand binding, we have now used NMR spectroscopy to characterize this enzyme in complex with a transition-state analog. The complex adopts a helix-bundle structure, as designed, but retains unprecedented flexibility on the millisecond timescale across its entire length. Moreover, pre–steady-state kinetics data show that binding occurs by an induced-fit mechanism on the same timescale as the enzymatic reaction, linking global conformational plasticity with efficient catalysis.
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Acknowledgements
We are grateful to P. Anikeeva for carrying out preliminary NMR analysis, J. Beld for analytical ultracentrifugation studies, R. Kissner for technical assistance with stopped-flow experiments, and K. Woycechowsky for critical reading of the manuscript. This work was supported by the Schweizerischer Nationalfonds and the ETH Zurich. We dedicate this paper to the memory of D. Koshland.
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K.P. and D.H designed research; K.P. and B.V. did NMR experiments; K.V. did biochemical experiments; K.P., K.V., B.V. and D.H. analyzed data; K.P., K.V. and D.H. wrote the paper.
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Supplementary Figures 1–4, Supplementary Tables 1–3, Supplementary Methods (PDF 1322 kb)
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Pervushin, K., Vamvaca, K., Vögeli, B. et al. Structure and dynamics of a molten globular enzyme. Nat Struct Mol Biol 14, 1202–1206 (2007). https://doi.org/10.1038/nsmb1325
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DOI: https://doi.org/10.1038/nsmb1325
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